Fig. 2

Reporter gene assays indicated a direct interaction of miR-142-5p with the ABL2 3′-UTR. Vectors containing the 3′-UTR of ABL2 were co-transfected with pre-miR-142-5p in two concentrations (10 nM, 25 nM). Reporter gene activities were measured 48 h after transfection. a Co-transfection of vector containing the ABL2 3′-UTR (WT) with miR-142-5p resulted in a suppression of relative reporter gene activity by 18% [10 nM] and 26% [25 nM]. b Introduction of mutation (MUT 1) into the predicted binding site at positions 3918–3921 bp of ABL2 3′-UTR led to a minor effect. c Combination of MUT 1 with mutations in the predicted binding site at positions 4247–4250 bp (MUT 2) abolished the inhibitory effect of the miRNA. d Predicted interactions of miR-142-5p with ABL2 3′-UTR is indicated and mutated bases for MUT 1 and MUT 2 are highlighted in gray. All reporter gene activities (n ≥ 12 in 3 independent experiments) were normalized to activities from cells transfected with respective 3′-UTR target sequence vectors and pre-miR negative control (median ± interquartile range). Activities are shown relative to empty control vector (c) identically transfected and normalized as 3′-UTR target sequence vectors. Mann–Whitney U-test; *p ≤ 0.05, ***p ≤ 0.001